title: Widespread dysregulation of peptide hormone release in mice lacking adaptor protein AP-3.
abstract: The regulated secretion of peptide hormones, neural peptides and many growth factors depends on their sorting into large dense core vesicles (LDCVs) capable of regulated exocytosis. LDCVs form at the trans-Golgi network, but the mechanisms that sort proteins to this regulated secretory pathway and the cytosolic machinery that produces LDCVs remain poorly understood. Recently, we used an RNAi screen to identify a role for heterotetrameric adaptor protein AP-3 in regulated secretion and in particular, LDCV formation. Indeed, mocha mice lacking AP-3 have a severe neurological and behavioral phenotype, but this has been attributed to a role for AP-3 in the endolysosomal rather than biosynthetic pathway. We therefore used mocha mice to determine whether loss of AP-3 also dysregulates peptide release in vivo. We find that adrenal chromaffin cells from mocha animals show increased constitutive exocytosis of both soluble cargo and LDCV membrane proteins, reducing the response to stimulation. We also observe increased basal release of both insulin and glucagon from pancreatic islet cells of mocha mice, suggesting a global disturbance in the release of peptide hormones. AP-3 exists as both ubiquitous and neuronal isoforms, but the analysis of mice lacking each of these isoforms individually and together shows that loss of both is required to reproduce the effect of the mocha mutation on the regulated pathway. In addition, we show that loss of the related adaptor protein AP-1 has a similar effect on regulated secretion but exacerbates the effect of AP-3 RNAi, suggesting distinct roles for the two adaptors in the regulated secretory pathway.
authors.affiliation: Graduate Program in Pharmaceutical Sciences and Pharmacogenomics, University of California San Francisco, San Francisco, California, United States of America ; Departments of Physiology and Neurology, University of California San Francisco, San Francisco, California, United States of America.
authors.forename: Daniel W
authors.initials: DW
authors.lastname: Sirkis
authors.forename: Robert H
authors.initials: RH
authors.lastname: Edwards
authors.forename: Cédric S
authors.initials: CS
authors.lastname: Asensio
mesh_terms: D033963:Adaptor Protein Complex 3 / Q000235:genetics* / Q000378:metabolism; D033966:Adaptor Protein Complex beta Subunits / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D003600:Cytosol / Q000378:metabolism*; D005089:Exocytosis / Q000235:genetics*; D005934:Glucagon / Q000235:genetics / Q000378:metabolism; D007328:Insulin / Q000235:genetics / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism; D008247:Lysosomes; D053858:Metabolic Networks and Pathways; D051379:Mice; D009474:Neurons / Q000378:metabolism; D036361:Peptide Hormones / Q000378:metabolism*; D034622:RNA Interference; D018808:Transcription Factor AP-1 / Q000235:genetics
chemical_list: D033963:Adaptor Protein Complex 3; D033966:Adaptor Protein Complex beta Subunits; C113910:Ap3d1 protein, mouse; D007328:Insulin; D036361:Peptide Hormones; D018808:Transcription Factor AP-1; D005934:Glucagon